• 한국수정란이식학회지
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• 제11권3호
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• pp.225-231
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• 1996

This study was conducted to investigate the effects of preincubation time, concentration and exposure time of sperm on in vitro fertilization of porcine follicular oocytes rnatured in in vitro. The results obtained are as follows ; 1. Effect of preincuhation time for porcine sperm capacitation on in vitro fertilization in medium with heparin was investigated. Normal fertilization rate was highest in 15 min(26.4%). However, there were no significant differences among preincuhation times of 5~90 min, 2. Normal fertilization rates of sperm concentrations were 17.0~26.5%, and normal fertilization rate from l$\times$ l05cell /ml concentration was also higher than those of other sperm concentration. 3. Normal fertilization rates of sperm exposure time of 4, 8, 12, 16 and 20 hours were 6.1, 20.8, 27.8, 25.0 and 26.7%, respectively. Normal fertilization rate from sperm exposure time of 12 hours was also higher than that of other sperm exposure times.

• Asian-Australasian Journal of Animal Sciences
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• 제9권4호
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• pp.371-374
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• 1996

Experiments were conducted to determine the effect of time intervals between collecting and use of cattle faeces as a source of micro-organisms in in vitro digestibility assays of forages. The results suggested that temperature conservation capacity by faeces depended on the size of the sample. There was no significant difference(p>0.05) between the first (T1 or 08:30 h) and second using time (T2 or 10:30 h). In vitro organic matter digestibility was significantly lower when faeces was used 5 h (T3 or 13:30 h) after collection. However, the organic matter digestibility determined at the second using time (T2) and third using time (T3) were highly correlated ($R^2=0.99$) with the first using time. It was concluded that faeces can be used as a source of microorganisms for in vitro digestibility assays of forages even 5 h after being voided.

• 한국가축번식학회지
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• 제21권2호
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• pp.103-109
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• 1997

The studies on the carried out to investigate to determine the optimum thawing temperature and equilibration time and 1 step straw method of frozen bovine embryos. The follicular oocytes were cultured in TCM-199 medium containing 10 IU/ml PMSG(Sigma, USA), 10 IU/ml hCG(Sigma, USA), 1 $\mu\textrm{g}$/ml $\beta$-estradiol(Sigma, USA) and 10% FCS for 24~48 hrs in incubator with 5% CO2 in air at 38.5$^{\circ}C$ and then matured oocytes were again cultured for 12~18 hrs with motile capacitated sperm by preincubation of heparin. The bovine embryos following dehydration by cryoprotective agents and various concentration of sucrose were directly plunged into liquid nitrogen and thawed in 3$0^{\circ}C$ water. Survival and in vitro developmental rate was defined as developmental rate on in vitro culture or FDA-test. The results are summarized as followes : 1. The equilibration time on in vitro developmental rates of bovine embryos was attained after short period of time(2.5~5 min.) in the freezing medium higher than long period of time (10~20 min.). 2. The temperature thawed at 3$0^{\circ}C$ after rapid freezing of bovine embryos resulted in a significantly higher in vitro developmental rate than did at 2$0^{\circ}C$ and 35$^{\circ}C$. 3. The thawing time on in vitro developmental rates of bovine embryos was attained after short period of time(1~5 min.) in the freezing mediuim higher than long period of time(10min.). 4. The in vitro developmental rates of bovine embryos after rapid frozen-thawing by 1 step straw method in the freezing medium added 1.5M, 2.0M glycerol, DMSO, propanediol and 0.25M, 0.50M, 0.75M, 1.00M sucrose were 12.5~19.4%, 10.0~15.6%, 9.1~13.8% and 6.7~12.9%, respectively.

• Reproductive and Developmental Biology
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• 제34권1호
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• pp.21-25
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• 2010

The purpose of this study was to investigate the effects of the collection time, co-culture and sperm penetration of canine oocytes on in vitro maturation and fertilization. The oocytes were cultured in TCM-199 media containing hormonal supplements (10% FCS, 10 IU/ml HCG, 10 IU/ml PMSG) at 5% $CO_2$, 95% air, $38^{\circ}C$. The in vitro maturation rate to MII stage of in vitro oocytes recovered from ovaries that collected at follicular, luteal and inactive phases of the reproductive phase for 44~72 hrs were 19.2%, 12.2%, and 6.0%, respectively. Follicular phases oocytes had a significantly higher in vitro maturation rate than oocytes collected at luteal and anestrus stage (p<0.05). The in vitro maturation rates to the MII stage of canine oocytes after 48 hrs of culture with glutathione, pyruvate, or glutathione + pyruvate were 12.5%, 10.7%, and 17.5%, respectively. This was higher than that in both alone or the combination of the two compared to the control group (19.0%). The sperm penetration rates of in vitro matured oocytes by fresh and frozen semen were 29/80 (36.3%) and 18/80 (22.5%), respectively. Although there are limited reports about canine oocytes co-culture and in vitro fertilization, our results on in vitro maturation is comparable to the results from other researches.

• 한국수정란이식학회지
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• 제12권1호
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• pp.27-36
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• 1997

The studies were carried out to investigate the effects of co-culture with cumulus cells and oviduct epithelial cells on the in vitro fertilization and cleavage rate of bovine follicular cocytes and to determine the optimum thawing temperature and equilibration time on in vitro developmental rate of frozen bovine embryos. The ovaries were obtained from slaughtered Korean native cows. The follicular oocytes were cultured in TGM-199 medium containing 10 IU /ml의 PM SG, 10 IU /ml의 hCG, ip g/ml의 $\beta$-estradiol and 10% FCS for 24~48 hrs in incubator with 5% $CO_2$ in air at 38.5$^{\circ}C$. The bovine embryos following dehydration by cryoprotective agents and a various concentration of sucrose were directly plunged into liquld nitrogen and thawed in 3$0^{\circ}C$ water. Survival rate was defined as developmental rate on in vitro culture or FDA-test. The results are sunanarized as followes :1. The in vitro fertilization and in vitro developmental rates of bovine oocytes co-cultured with cumulus cells in TCM499 medium were 75.0~76.8% and 17.3~27.6%, respect-ively. And in-vitro fertilization rates of cumulus-enclosed oocytes(55.4%)were significantly(p<0.05) higher than cumulus-denuded oocytes (23.1%). 2. The in vitro fertilization and in vitro developmental rates of bovine oocytes co-cultured with l$\times$ l04cells /ml, 1 x l06cells /ml, lx l08cells /ml and 1 x l015cells /ml oviduct epithelial cells in TCM-199 medium were 74.5~77.8% and 15.7~21.20 respectively.3. The in-vitro fertilization and in vitro developmental rates of bovine oocytes cocultured in '1CM-199 media containing PMSG, hCG, PMSG+hCG. PMSG+$\beta$-estradiol, hCG+$\beta$-estradiol 0 to 40 hrs after insemination were 74.0~77.4% and l8.9~23.l%, re-spectiv ely.4.The survival rates of bovine embryos thawed after rapid freezing in the freezing medium containing a various concentration of sucrose added 1.5M and 2.OM glycerol,DMSO and propanediol were 23.5~31.4% and 20.6~34.l%, respectively. 5. The temperature thawed at 3$0^{\circ}C$ after rapid freezing of bovine embryos resulted in a significantly higher embryos survival rate than did at 2$0^{\circ}C$ and 35$^{\circ}C$.6. The equilibration time on the survival rates of bovine embryos was attained after short period of time(2.5~5 min.) in the freezing medium higher than long period of time (10~20min.). (Key words : bovine embryos, co-culture, freezing, in vitro development)

• 한국동물생명공학회지
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• 제37권4호
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• pp.285-291
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• 2022

In 1951, Colin Russell Austin and Min Chueh Chang identified "capacitation", a special process involving ejaculated spermatozoa in the female reproductive tract. Capacitation is a phenomenon that occurs in vivo, but almost all knowledge of capacitation has been obtained from in vitro studies. Therefore, numerous trials have been performed to establish in vitro capacitation methods for various studies on reproduction. Although a series of studies have been conducted to develop an optimal protocol for inducing capacitation, most have focused on identifying the appropriate chemical compounds to induce the capacitation of boar spermatozoa in vitro. Therefore, the purpose of this study was to identify the optimal incubation time for inducing capacitation in vitro. Duroc semen was incubated for various periods (60, 90, and 120 min) to induce capacitation. Sperm function (sperm motility, motion kinematic parameters, and capacitation status) was evaluated. The results showed that total sperm motility, rapid sperm motility, progressive sperm motility, curvilinear velocity, and average path velocity significantly decreased in a time-dependent manner. However, the capacitation status did not show any significant changes. Taken together, these results indicate that an incubation time of more than 60 min suppresses sperm motility and motion kinematic parameters. Therefore, we suggest that 60 min may be the best incubation time to induce capacitation without negative effects on sperm motility and motion kinematics in boar spermatozoa in vitro.

• 한국수정란이식학회지
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• 제15권1호
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• pp.33-38
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• 2000

This study was undertaken in an effort to select the sire bull in Hanwoo through in vitro fertilization of proven bull semen. It was used for in vitro fertilization that of the 20 proven bull semen with follicular oocytes derived from slaughterhouse ovaries of Hanwoo. The stage of maturation on the time course of bovine cumulus-enclosed oocytes incubated for 24 hours was found the highest(96.4%) than hose of other maturationi time. In vitro fertilization rate of bovine oocytes with proven bull sperm showed from 61.5 to 88.9%. Polyspermy of in vitro fertilized oocytes according to proven bulls were the highest KP 491(61.5%) nothing but KP 486, KP 491 and KP 497.

• 한국가축번식학회지
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• 제15권2호
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• pp.141-147
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• 1991

This stduy was carried out in order to investigate the effects of cryoprotective concentration and equilibration time on survival rate of ultrarapidly frozen in vitro fertilized bovine embryos. In vitro fertilized bovine embryos, following dehydration by cryoprotective agents and sucorese were directly plunged into liquid nitrogen and thawed in 38$^{\circ}C$ water. Survival rate was defined by development rate to the morula and blaqstocyst stage after in vitro culture of by FDA test. The results are summarized as follows : 1. The survival rates of in vitro fertilized bovine embryos after ultrarapid frozen-thawing in the freezing medium of 0.25M sucroese added 2.0M, 2.5M, 3.0M, 3.5M, 4.0M glycerol were 75.0%, 72.0%, 67.6%, 44.8% and 18.3% respectively. 2. The survival rates of in vitro fertilized bovine embryos after ultrarapid frozen-thawing in the freezing medium of 0.25M sucrose added 2.0M, 2.5M, 3.0M, 3.5M, 4.0M DMSO were 64.0%, 66.7%, 70.8%, 52.7% and 18.6, respectively. 3. The survival rates of in vitro fertilized bovine embryos after ultrarapid frozen-thawing in the freezing medium of 0.25M sucrose added 2.0M, 2.5M, 3.0M, 3.5M, 4.0M propanediol were 68.4%, 64.9%, 63.2%, 62.2% and 34.7%, respectively. 4. The survival rates of in vitro fertilized bovine embryos after ultrarapid frozen-thawing in the freezing medium of 2.50M glycerol added 0.1M, 0.25M, 0.5M, 0.75M, sucrose were 60.5%, 72.2%, 70.1% and 54.9%, respectively. The survival rate of in vitro fertilized embryos after ultrarapid frozen-thawing in the freezing medium of 2.5M glycerol added 0.25M sucrose were higher than concentration of 0.10M, 0.50M and 0.75M sucrose. 5. The equilibration time on the survival rate of in vitro fertilized bovine embryos was attained after short period of time(2.5~5min.) in the freezing medium added 0.25M sucrose and 3.0M DMSO higher than long period time(1~20min.).

• Journal of Pharmaceutical Investigation
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• 제32권4호
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• pp.321-325
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• 2002

IVIVC (In vitro/in vivo correlation) is useful for predicting in vivo results from in vitro data. The aim of this study was to develop IVIVC of sustained release diltiazem. For this purpose, three types of diltiazem tablets with different in vitro dissolution rates were prepared. An in vitro dissolution testing method comprising of paddle apparatus, 50 rpm, water as dissolution medium was developed. Under these condition, we demonstrated that AUCinf could be predicted by evaluating $d_{70%}$ (time dissolved 70%) in vitro since the in vivo AUCinf was correlated with the in vitro $d_{70%}$ (r=-0.9981).

• Journal of Pharmaceutical Investigation
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• 제23권4호
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• pp.217-223
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• 1993

The combined products of antacid and anti-ulcer agent were prepared with antacid composed of aluminium hydroxide dried gel, magnesium hydroxide and simethicone with a ratio of 1:1:0.1 (M) and anti-ulcer agent, aceglutamide aluminium (AGA). The efficacy of antacid was evaluated in vitro with Fuchs, Johnson-Duncan and Rosset-Rice methods and in vivo using an aspiration method in rat. The addition of anti-ulcer agent did not affect the neutralizing capacity of M significantly. The combined products with the M/AGA ratios of 2.3:1 and 3.4:1 produced the maximum pH of $4.0{\sim}5.8$ and the duration time of $64{\sim}137$ min in vitro test. The in vivo neutralizing test in rats showed the rapid increase of gastric pH up to 3.5 within 30 min and the gastric pH of $4{\sim}6$ was kept for 5 hr.